Advanced

Lentiviral-mediated gene transfer into haematopoietic stem cells

Woods, N B LU ; Mikkola, H ; Nilsson, E LU ; Olsson, K LU ; Trono, D and Karlsson, S (2001) In Journal of Internal Medicine 249(4). p.339-343
Abstract

OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.

DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.

RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the... (More)

OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.

DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.

RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the secondary recipients 6 weeks posttransplantation. Human haematopoietic progenitor colonies were grown from both primary and secondary recipients. Over 50% of the haematopoietic colonies in these recipients were positive for the GFP transgene by PCR. Following inverse PCR, amplified fragments were sequenced and integration of the vector into human genomic DNA was demonstrated. Several vectors containing different internal promoters were tested in NOD/SCID mice that had been transplanted with transduced CD34+ and CD34+/CD38- cells. The elongation factor-1alpha (EF-1alpha) promoter gave the highest level of expression, both in the myeloid and lymphoid progeny of the engrafting cells.

CONCLUSIONS: These data collectively indicate that candidate human HSCs can be efficiently transduced with lentiviral vectors and that the transgene is highly expressed in their progeny cells.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Antigens, CD34, Gene Transfer Techniques, Genetic Vectors, Hematopoietic Stem Cells, Humans, Lentivirus, Mice, Mice, SCID, Promoter Regions, Genetic, Transduction, Genetic
in
Journal of Internal Medicine
volume
249
issue
4
pages
5 pages
publisher
Wiley-Blackwell Publishing Ltd
external identifiers
  • pmid:11298854
  • scopus:0035731632
ISSN
0954-6820
DOI
10.1046/j.1365-2796.2001.00806.x
language
English
LU publication?
yes
id
a731b6d7-23d7-4aa5-bf64-1486ed9a8daa
date added to LUP
2019-11-25 13:51:03
date last changed
2020-01-16 04:07:48
@article{a731b6d7-23d7-4aa5-bf64-1486ed9a8daa,
  abstract     = {<p>OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.</p><p>DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.</p><p>RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the secondary recipients 6 weeks posttransplantation. Human haematopoietic progenitor colonies were grown from both primary and secondary recipients. Over 50% of the haematopoietic colonies in these recipients were positive for the GFP transgene by PCR. Following inverse PCR, amplified fragments were sequenced and integration of the vector into human genomic DNA was demonstrated. Several vectors containing different internal promoters were tested in NOD/SCID mice that had been transplanted with transduced CD34+ and CD34+/CD38- cells. The elongation factor-1alpha (EF-1alpha) promoter gave the highest level of expression, both in the myeloid and lymphoid progeny of the engrafting cells.</p><p>CONCLUSIONS: These data collectively indicate that candidate human HSCs can be efficiently transduced with lentiviral vectors and that the transgene is highly expressed in their progeny cells.</p>},
  author       = {Woods, N B and Mikkola, H and Nilsson, E and Olsson, K and Trono, D and Karlsson, S},
  issn         = {0954-6820},
  language     = {eng},
  number       = {4},
  pages        = {339--343},
  publisher    = {Wiley-Blackwell Publishing Ltd},
  series       = {Journal of Internal Medicine},
  title        = {Lentiviral-mediated gene transfer into haematopoietic stem cells},
  url          = {http://dx.doi.org/10.1046/j.1365-2796.2001.00806.x},
  doi          = {10.1046/j.1365-2796.2001.00806.x},
  volume       = {249},
  year         = {2001},
}